Biomedical subjects

R A Davis

Publications and source records attributed to R A Davis.

At least 37 records · Page 2Linked to original sources

PubMed1997

Coordinate regulation of lipogenesis, the assembly and secretion of apolipoprotein B-containing lipoproteins by sterol response element binding protein 1.

Stable plasmid-driven expression of the liver-specific gene product cholesterol 7alpha-hydroxylase (7alpha-hydroxylase) was used to alter the cellular content of transcriptionally active sterol response element binding protein 1 (SREBP1). As a result of stable expression of 7alpha-hydroxylase, individual single cell clones expressed varying amounts of mature SREBP1 protein. These single cell clones provided an opportunity to identify SREBP1-regulated genes that may influence the assembly and secretion of apoB-containing lipoproteins. Our results show that in McArdle rat hepatoma cells, which normally do not express 7alpha-hydroxylase, plasmid-driven expression of 7alpha-hydroxylase results in the following: 1) a linear relationship between (i) the cellular content of mature SREBP1 and 7alpha-hydroxylase protein, (ii) the relative expression of 7alpha-hydroxylase mRNA and the mRNA's encoding the enzymes regulating fatty acid, i.e. acetyl-CoA carboxylase and sterol synthesis, i.e. HMG-CoA reductase, (iii) the relative expression of 7alpha-hydroxylase mRNA and microsomal triglyceride transfer protein mRNA, a gene product that is essential for the assembly and secretion of apoB-containing lipoproteins; 2) increased synthesis of all lipoprotein lipids (cholesterol, cholesterol esters, triglycerides, and phospholipids); and 3) increased secretion of apoB100 without any change in apoB mRNA. Cells expressing 7alpha-hydroxylase contained significantly less cholesterol (both free and esterified). The increased cellular content of mature SREBP1 and increased secretion of apoB100 were concomitantly reversed by 25-hydroxycholesterol, suggesting that the content of mature SREBP1, known to be decreased by 25-hydroxycholesterol, mediates the changes in the lipoprotein assembly and secretion pathway that are caused by 7alpha-hydroxylase. These data suggest that several steps in the assembly and secretion of apoB-containing lipoproteins by McArdle hepatoma cells may be coordinately linked through the cellular content of mature SREBP1.

Acetyl-CoA Carboxylase↗

PubMed1997

Expression of human cholesterol 7alpha-hydroxylase in atherosclerosis-susceptible mice via adenovirus infection.

Adenovirus is a vector for the delivery of genes mainly to the liver. Short-term (approximately 3 days) studies using adenovirus transfection have provided valuable insights into how genes can complement normal and pathological phenotypes. When atherosclerosis-susceptible C57BL/6 mice were infected with an adenovirus vector containing the human 7alpha-hydroxylate cDNA (AV17h1) and fed on a chow diet, human 7alpha-hydroxylase mRNA and enzyme activity doubled compared with that in mice infected with an adenovirus vector (AV1Null) alone. In AV17h1-infected mice fed on a high fat cholic acid (HFCA) diet, mRNA expression and activity of both the endogenous and adenovirus (human) 7alpha-hydroxylase were repressed. AV17h1-infected mice fed on a HFCA diet and killed at mid-light had increased 7alpha-hydroxylase activity and mRNA compared with mice killed at mid-dark. Since expression of AV17h1 is driven by a constitutive Rous sarcoma virus promoter, the repression of human 7alpha-hydroxylase by the HFCA diet was unexpected. In spite of this post-transcriptional repression by the HFCA diet, AV17h1-infected mice expressed the human 7alpha-hydroxylase mRNA, causing its enzyme activity to be 3-fold greater than in AV1Null-infected mice. In AV17h1-infected mice, the 7alpha-hydroxylase enzyme activity varied as a linear function of human mRNA abundance. In conclusion, the accumulation of apolipoprotein B-containing lipoproteins in plasma of C57BL/6 mice fed on the HFCA diet was not reduced by longer-term (2 weeks) 7alpha-hydroxylase expression, probably because of its diminished expression caused by the diet and hepatic inflammation from the adenovirus infection. These results may suggest that adenovirus is effective in promoting longer-term (2 weeks) expression of 7alpha-hydroxylase.

Adenoviridae Infections↗

PubMed1997

Transcriptional induction of cholesterol 7alpha-hydroxylase by dexamethasone in L35 hepatoma cells requires sulfhydryl reducing agents.

It is known that hepatic levels of reduced glutathione correlate with the activity of the liver-specific enzyme cholesterol-7alpha-hydroxylase. We examined the possibility that sulfhydryl reducing agents activate transcription of cholesterol 7alpha-hydroxylase. Adding dithiothreitol (DTT, 1 mM) and dexamethasone to L35 hepatoma cells increased the content of 7alpha-hydroxylase mRNA 3-fold above the levels observed with dexamethasone alone. Without dexamethasone, DTT had no affect. The addition of reduced glutathione to L35 cells demonstrated a similar potentiation of expression dependent on dexamethasone. Nuclear run-on assays showed that in the presence of both dexamethasone and DTT, the transcription of the 7alpha-hydroxylase gene was clearly increased. In contrast, by itself, dexamethasone did not cause a detectable increase in the transcription of the 7alpha-hydroxylase gene. Dexamethasone and DTT did not affect the transcription of beta-actin, suggesting a selective induction of the 7alpha-hydroxylase gene. DTT reversed repression of 7alpha-hydroxylase expression by insulin but not the repression by phorbol ester. Our data show for the first time that the sulfhydryl redox potential of the hepatocyte (i.e. level of reduced glutathione) has a marked influence on the transcription and expression of the liver-specific gene 7alpha-hydroxylase.

Animals↗

PubMed1997

National incidence of smoking and misclassification among the U.S. married female population.

Because of a lack of representative data on smoking status misclassification among U.S. married females, a two-part study was conducted. Part I was conducted to obtain nationally representative estimates of the percentage of U.S. women who report themselves to be current, former, and never smokers, to determine the concordance of smoking habits among spouse pairs, and to establish field quotas and probability weightings for Part II. Part II was conducted to determine smoker misclassification rates using salivary cotinine as an indication of active smoking. Part I, conducted in January 25-29, 1992, utilized random-digit dialing telephone interviewing throughout the 48 contiguous United States. Part II, conducted from February 19, 1992 to March 7, 1992, was a mall-intercept study in nine geographically disperse U.S. cities and it involved interviewing and saliva collection. Among married U.S. women, 25% reported they were current smokers, 22% reported they were former smokers, and 53% reported they were never smokers. Using a cotinine concentration of either > 35 ng/ml or > 106 ng/ml to indicate regular smoking, 3.61% and 2.55% of regular smokers, respectively, reported themselves to be never smokers. The concordance ratio, an important parameter in correcting for non-differential misclassification bias, was found to be 5.52. In addition, an indication of substantial differential misclassification was found between exposed and unexposed populations. This type of misclassification bias has previously not been accounted for in the adjustment of epidemiology-based risk assessments of tobacco smoke exposure and lung cancer. Taken together, these data suggest that misclassification bias alone is likely to explain any lung cancer risk elevation observed in the U.S. epidemiology of environmental tobacco smoke exposure among nonsmoking women.

Cotinine↗

PubMed1997

Evolution of processes and regulators of lipoprotein synthesis: from birds to mammals.

Mammalian lipoproteins are synthesized in the liver and secreted into the blood plasma where they are targeted to specific tissues. Through specific cell surface receptors, hepatic lipoproteins are taken up and their lipid contents are then used for anabolic and energy requirements. Because of the well-established role that plasma lipoproteins play as risk factors for the development of cardiovascular disease, a great amount of attention has been directed toward understanding their metabolism and biosynthesis. The major focus of this report is to review the evolution of gene products that are essential in regulating, synthesizing, assembling and secreting the lipid and protein components of lipoproteins. Using the primordial vitellogenin lipoprotein system as the paradigm, I show how metabolic signals derived from the sterol biosynthetic pathway provide a coordinate regulation of genes necessary to assemble and secrete mammalian apolipoprotein B-containing lipoproteins. In lower species, estrogen induces the expression of genes required for both vitellogenin synthesis and its tissue targeting (the vitellogenin receptor). This coordinate induction provides lipid to the ovaries for egg development. In mammals, a sterol-derived metabolic signal regulates the expression of genes required for lipoprotein synthesis and for the LDL receptor. The signal is not sex linked, providing an adaptive advantage to changes in nutritional status.

Animals↗

PubMed1997

Clinical evaluation of Acuvue contact lenses with UV blocking characteristics.

BACKGROUND: In response to increasing scientific evidence which indicates that ultraviolet radiation (UVR) is a potential threat to ocular health, Acuvue contact lenses (Vistakon, Johnson & Johnson Vision Products Inc., Jacksonville, Florida) have been developed which incorporate an ultraviolet (UV) blocker within the lens polymer. Data are presented for the first clinical evaluation of Acuvue lenses with UV blocking characteristics. METHOD: A double-masked, multicenter, prospective clinical trial involving 94 subjects was conducted. The study followed a randomized, parallel group design and consisted of 3 months of daily wear with two-weekly lens replacement. Two thirds of the subjects (61) wore the test lenses (Acuvue with UV blocker) and the remaining subjects (33) wore conventional Acuvue lenses (without UV blocker). RESULTS: Biomicroscopic evaluations indicated that the performance of the test and control lenses was clinically similar. No clinically relevant differences between the test and control lenses were noted in the subjective assessments of vision, comfort, or handling. In addition, no differences were shown for surface deposition, lens durability, visual acuity, and subjective symptoms. CONCLUSION: The study findings indicate that the addition of a UV blocker to Acuvue contact lenses has been achieved without affecting daily wear clinical performance. Because there is increasing evidence to suggest that the ocular tissues may be damaged by UVR, it is prudent for eye care practitioners to prescribe contact lenses that offer the benefits of both regular replacement and UV protection.

Adolescent↗

PubMed1997

Cholesterol 7alpha-hydroxylase influences the expression of hepatic apoA-I in two inbred mouse strains displaying different susceptibilities to atherosclerosis and in hepatoma cells.

C57BL/6 mice are susceptible to diet-induced atherosclerosis, whereas BALB/c mice are resistant. The susceptibility of C57BL/6 mice has been linked to decreased plasma HDL cholesterol in response to a diet containing fat, cholesterol, and cholic acid. Feeding C57BL/6 mice a diet consisting of fat and cholesterol, but no cholic acid, increased plasma high density lipoprotein (HDL) cholesterol. The increase in HDL was associated with increases in both plasma apolipoprotein (apo)A-I and hepatic apoA-I mRNA. Supplementation of the cholesterol-rich diet with cholic acid inhibited the stimulatory effect of cholesterol on hepatic apoA-I mRNA expression, resulting in similar hepatic apoA-I mRNA levels compared to chow-fed mice. Atherosclerosis-resistant BALB/c mice were also resistant to diet-induced changes in plasma HDL, apoA-I, and hepatic apoA-I mRNA levels. Previous studies showed that the diets changed both the activity and mRNA encoding the liver specific enzyme 7alpha-hydroxylase (1993.J. Lipid Res. 34: 923-931). In both strains of mice, hepatic expression of apoA-I and 7alpha-hydroxylase mRNA varied in parallel. Whereas susceptible C57BL/6 mice also showed a significant correlation between HDL cholesterol and expression of 7alpha-hydroxylase, no such correlation was observed in BALB/c mice, suggesting that genetic differences in HDL metabolism, not hepatic apoA-I synthesis, are responsible for the strain specific differences in plasma HDL levels. The finding that lecithin: cholesterol acyltransferase (LCAT) activity was significantly decreased in C57BL/6 mice, but not in BALB/ c mice fed the atherogenic diet, further supports this conclusion. Additional studies show that McArdle hepatoma cells stably expressing plasmid-derived rat 7alpha-hydroxylase recapitulated the parallel linear relationship between 7alpha-hydroxylase and apoA-I mRNA expression observed in both strains of mice. These data link hepatic apoA-I mRNA expression to hepatic cholesterol/bile acid metabolism.

Animals↗

PubMed1996

Human urine mutagenicity study comparing cigarettes which burn or primarily heat tobacco.

Cigarette smokers have been reported to void urine which is more mutagenic, as measured in the Ames assay, than urine voided by non-smokers. Condensate from the mainstream smoke of a cigarette which primarily heats tobacco (test cigarette) has shown significantly reduced mutagenicity in a battery of in vitro genotoxicity assays compared with tobacco-burning cigarettes. The objective of this study was to determine whether the reduction in mutagenic activity observed in the in vitro assays would be reflected in the urine of smokers of the test cigarette. Twenty smokers were enrolled in a 4-week crossover study, with each smoker consuming test cigarettes ad libitum for a week and their usual brand of tobacco-burning cigarettes the other 3 weeks. Diet was strictly controlled throughout the study, and broiled and pan-fried meat was not served to minimize ingestion of mutagenic protein pyrolysis products. There was no statistically significant difference (p = 0.06) in consumption of tobacco-heating and tobacco-burning cigarettes. There were no statistically significant differences (p = 0.22) in salivary cotinine concentrations for smokers when smoking either tobacco-burning or tobacco-heating cigarettes. Urinary nicotine (ng/mg creatinine) was not different (p = 0.31) for smokers when smoking either tobacco-burning or tobacco-heating cigarettes. Urinary cotinine (ng/mg creatinine) was 32% lower (p = 0.0004) when smoking tobacco-heating cigarettes as compared with smoking tobacco-burning cigarettes. Twenty-four-hour urine samples were collected twice weekly, concentrated using XAD-2 resin and tested in Ames strains TA98 and YG1024 with metabolic activation. Tobacco-burning cigarette smokers experienced a 79% reduction in urinary mutagenicity as measured in strain YG1024 and a 72% reduction as measured in strain TA98 during the week that they smoked the tobacco-heating cigarette while maintaining a fixed dietary regimen. The results of this study indicate that smokers of tobacco-heating cigarettes void urine which is significantly less mutagenic than urine voided by smokers of tobacco-burning cigarettes.

Cotinine↗

PubMed1996

Thermodynamic stabilization of nucleotide binding to thymidylate synthase by a potent benzoquinazoline folate analogue inhibitor.

The stabilization of dUMP, FdUMP, and dGMP binding to Escherichia coli thymidylate synthase (TS) in the presence and absence of a folate analogue inhibitor of TS, 1843U, was determined by differential scanning calorimetry. When the enzyme is thermally unfolded in the presence of dUMP, two separate temperature transitions are evident, although only one binding site/dimer was detected in equilibrium dialysis experiments. In the absence of dUMP, TS shows a major peak of unfolding at 45 degrees C with a shoulder at 47 degrees C. In the presence of increasing amounts of dUMP progressive changes in the size of each peak occur, each associated with a higher temperature of unfolding. At a ratio of dUMP/TS of 100, a major peak predominates with an unfolding temperature (Td) of 60 degrees C. FdUMP shows a similar profile, while dGMP does not alter the Td of the enzyme since dGMP alone does not bind to TS. Despite the fact that 1843U binds tightly to TS in the absence of nucleotide ligands [Dev, I. K., Dallas, W.S., Ferone, R., Hanlon, H., McKee, D.D., & Yates, B. B. (1994) J.Biol. Chem. 269, 1873-1882], it exhibits only a small effect on the Td profile of TS. However, when 1843U is present, in addition to the nucleotides (dUMP, FdUMP, or dGMP), a Td of 72 degrees C is achieved and the enthalpy of unfolding is increased by one-third. The stabilizing effect of substrate binding to TS by 1843U examined by thermodynamic parameters can be attributed to the considerable extra amount of free energy released on formation of the ternary complex of TS-1843U-nucleotide. The tightness of this complex is due to the stacking energy that results from Van der Waals contacts between the nucleotide purine or pyrimidine ring and the benzoquinazoline ring of 1843U [Weichsel, A., Montfort, W. R., Cieśla, J., & Maley, F. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 3493-3497], which induces a local conformational change in the protein. This conformational change is associated with a significant positive entropy change, which suggests that water is expelled from the active site region.

Calorimetry, Differential Scanning↗

PubMed1996

Assessing the sensory role of nicotine in cigarette smoking.

Thirty-two subjects were tested in five double-blind sessions (16 subjects in the morning following overnight smoking abstention, and 16 in the afternoon following ad-lib smoking). In each session, subjects smoked one of five experimental (EX) cigarettes having the following FTC nicotine/"tar' yields in mg: 0.08/8.5, 0.17/9.1, 0.37/9.8, 0.48/9.8, and 0.74/10.4. In a sixth session, subjects smoked a 0.71/8.6 commercial "light' (CL) cigarette that was their usual brand. Before and after smoking, subjects subjectively rated their desire to smoke a cigarette of their usual brand and had blood samples drawn. Following smoking, subjects rated the cigarette on a variety of sensory dimensions; they also rated smoking satisfaction. Analysis of variance indicated that nicotine played an important sensory role for a variety of dimensions related to cigarette taste and sensory impact but not perceived draw. Principal-components analyses indicated that sensory factors were at least as important as nicotine pharmacology (indirectly indexed by the pre-to post-smoking rise in blood nicotine concentration) when considering smoking's overall effects on satisfaction, product acceptance, and reduction in desire to smoke.

Adult↗

PubMed1996

A long-term outcome study of 170 surgically treated patients with compressive cervical radiculopathy.

BACKGROUND: There is a general impression that publications in neuro-surgical journals have comprehensively evaluated the relative merits of various operative techniques with long-term follow-up in patients treated for compressive cervical radiculopathy. However, in some 20,000 neurosurgical papers over the past 50 years, only 74 (0.3%) have addressed this common surgical problem. METHODS: Long-term outcome was assessed by the standardized Functional Economic Outcome Rating Scale of Prolo. Information was obtained primarily by telephone interviews and office visits. No patients were operated upon without a confirmatory neuroradiologic study. Single level soft discs were removed through a posterior-lateral foraminotomy with hemilaminectomy or laminectomy the root was decompressed by the same technique with hard discs. RESULTS: One hundred seventy patients were operated on from 1959-91 with a 96% follow-up. The mean follow-up period was 15 years. Patients who had sedentary occupations and housewives, had statistically higher Economic Prolo scores (p < 0.001) than those who did strenuous work. Of 10 patients with a total Prolo score of 5 or less, seven did strenuous work and had Workers' Compensation claims; the remaining had legal claims or were at psychologic risk for operation. In 86% of patients outcome was good; defined as a Prolo score of 8 in 5%, 9 in 38%, and 10 in 43%. Of 10 recurrences (6%), seven occurred within 3 years after operation and were treated by discectomy and anterior cervical fusion. There were two patients with postoperative deltoid motor weakness who recovered within 1 year. CONCLUSIONS: Although outcome studies must have subjective criteria, the Prolo Scale is more objective and quantitative than currently used methods. The posterior approach to disc lesions causing compressive cervical radiculopathy should be compared to other techniques using the Prolo Outcome Scale.

Adult↗

PubMed1996

Psychophysiological and subjective effects of cigarettes having varying nicotine yields but relatively constant "tar' yields.

Thirty-two subjects were tested in five double-blind sessions-16 subjects in the morning (a.m.) following overnight smoking abstention, and 16 in the afternoon (p.m.) following ad lib smoking. In each session, subjects smoked 1 of 5 cigarettes having the following FTC nicotine/'tar' yields in mg: 0.08/8.5, 0.17/9.1, 0.37/9.8, 0.48/9.8, and 0.74/10.4. On a pre- to postsmoking basis, blood nicotine and heart rate increased with nicotine yield. The effect of nicotine yield on changes in self-rated anxiety was an inverted-U function, but this effect was possibly confounded by baseline differences. The following effects on EEG spectral-band magnitude were also obtained: (1) nicotine yields > or = 0.17 mg decreased delta; (2) nicotine yields of 0.37 and 0.48 mg decreased theta in a.m. subjects; (3) nicotine yields > or = 0.37 mg decreased alpha, the effect being greater in a.m. subjects; (4) no effect of yield on beta 1 was obtained; (5) nicotine yields of 0.48 and 0.74 mg increased the Cz-minus-T5 differential in beta 2. Pre- to postsmoking changes in this measure of beta 2 were not correlated with either blood nicotine or anxiety.

Adolescent↗

PubMed1996

Pentobarbital alters the response of cerebrospinal fluid pressure evoked by two provocative tests in rats.

The intracerbroventricular (ICV) route of administration is a commonly used pharmacologic procedure in rats. However, little attention is given to changes in cerebrospinal fluid pressure (CSFp) when the ventricular system is perturbed, especially in conscious animals. This communication examined CSFp in pentobarbital-anesthetized rats subjected to two pertubations where CSFp had been previously measured in conscious rats. Pentobarbital treatment resulted in marked, qualitative differences after both protocols. Whereas conscious rats responded to a 10-min ICV infusion (8 microliters/min) by a delayed, secondary increase in CSFp, this effect was completely eliminated (p < 0.001) by pentobarbital. Also, whereas conscious rats could maintain CSFp at normal values during an hour of 45 degrees head-down tilt, pentobarbital-treated rats could not. Anesthetized rats displayed an immediate fall in CSFp to near zero values (p < 0.001), followed by a rapid recovery (p < 0.001) after tilting. Such differences in CSF dynamics raise important issues. For example, what mechanism(s) underlie the changes in CSF dynamics? Also, are these differences in conscious and anesthetized rats potential variables for drug distribution when the ICV route is used?

Anesthesia↗

PubMed1996

Rat hepatoma L35 cells, a liver-differentiated cell line, display resistance to bile acid repression of cholesterol 7 alpha-hydroxylase.

A stable hepatoma cell line (L35 cells) showing an activation of the cholesterol 7 alpha-hydroxylase gene (CYP7) that had been silent in the parental hepatoma cell line (H35 cells) was used to examine the influence of bile acids on its gene expression and activity. L35 cells were found to concentrate taurocholate from the culture medium, without any significant effect on the expression of 7 alpha-hydroxylase. At physiologic levels (up to 100 microM), CYP7 mRNA expression was not repressed by any bile acid. At supra-physiologic levels (1 mM), the more hydrophobic dihydroxy bile acids, taurodeoxycholate and taurochenodeoxycholate, decreased CYP7 mRNA without decreasing the relative abundance of beta-actin mRNA. Similar results were obtained by culturing cells with sodium dodecylsulfate (50 microM). The medium of L35 cells treated with either taurochenodeoxycholate (1 mM), taurodeoxycholate (1 mM), or sodium dodecylsulfate (50 microM) contained significantly greater activities of two cytosolic enzymes, lactate dehydrogenase and phosphoglucose isomerase, indicating a cytotoxic response. Activation of protein kinase C by phorbol esters decreased the expression of 7 alpha-hydroxylase mRNA without evidence of cytotoxicity; therefore, the inability of L35 cells to show bile acid repression cannot be ascribed to a lack of an effect by this secondary messenger system. In addition, insulin decreased and dexamethasone increased 7 alpha-hydroxylase mRNA without increasing the release of the cytoplasmic enzyme markers. The combined data suggest that L35 cells are resistant to repression of CYP7 gene expression by bile acids, but display physiologic expression to hormones and protein kinase C activation.

Animals↗

PubMed1996

Translocation of apolipoprotein B across the endoplasmic reticulum is blocked in abetalipoproteinemia.

Abetalipoproteinemia (ABL) is an autosomal recessive disease characterized by the inability of the liver and intestine to secrete apolipoprotein B (apoB). Mutations in the microsomal triglyceride transfer protein (MTP) gene, but not the apoB gene, are responsible for the ABL phenotype. It is not clear how loss of MTP in ABL patients leads to a complete, but specific, block in the secretion of apoB. It is to this question that our work is directed. In cultured cells lacking MTP, translocation of apoB is completely arrested, leading to the hypothesis that apoB requires MTP in order to completely enter the lumen of the endoplasmic reticulum, the site of lipoprotein assembly. We examined this hypothesis by determining the presence in plasma of distinct N-terminal apoB peptides, produced exclusively from translocation arrested apoB, in the plasma of six ABL patients and six normal subjects. The data show that N-terminal apoB peptides are present in the plasma of six ABL patients, whereas intact apoB-100 was barely detectable. Moreover, the plasma of all six ABL patients displayed a 2000-fold increase in the amount of an 85 kDa N-terminal apoB peptide relative to apoB-100. These data provide the first in vivo data supporting the essential role that MTP plays in apoB translocation. In normal humans, varied expression of MTP may be responsible for the post-transcriptional regulation of apoB secretion.

Abetalipoproteinemia↗

PubMed1995

In HepG2 cells, translocation, not degradation, determines the fate of the de novo synthesized apolipoprotein B.

Previous studies show that translocation and degradation of apolipoprotein B (apoB), two processes occurring on or within the endoplasmic reticulum, determine how much de novo synthesized apoB is secreted. We determined which of these processes regulates the intracellular fate of apoB by examining whether degradation determines how much apoB is translocated or if translocation determines how much apoB is degraded. HepG2 cells, treated with the cysteine active site protease inhibitor ALLN, previously shown to block the degradation of translocation-arrested apoB in Chinese hamster ovary cells (Du, E., Kurth, J., Wang, S.-L., Humiston, P., and Davis, R.A. (1994) J. Biol. Chem. 269, 24169-24176), showed a 10-fold increase in the accumulation of de novo synthesized [35S]methionine-labeled apoB. The majority (80%) of the apoB accumulated in response to ALLN was in the microsomal fraction. In contrast, ALLN did not effect apoB secretion. Since ALLN did not effect the intracellular accumulation of [35S]methionine-labeled albumin and other proteins (trichloroacetic acid-precipitable [35S]methionine-labeled proteins), its effect on apoB was specific. Pulse-chase studies showed that ALLN dramatically reduced the first-order rate of removal of [35S]methionine-labeled apoB from the cell but did not effect its rate of secretion. The finding that ALLN caused the intracellular accumulation of incompletely translated chains of apoB suggests that at least some of the degradation occurs at the ribosomal level. Moreover, 85% of the apoB that accumulated in isolated microsomes in response to ALLN was accessible to exogenous trypsin, indicating this pool of apoB was incompletely translocated. The combined data suggest that translocation, not degradation, determines the intracellular fate of de novo synthesized apoB.

Apolipoprotein B-100↗

PubMed1995

Dynamics of MASH1 expression in vitro and in vivo suggest a non-stem cell site of MASH1 action in the olfactory receptor neuron lineage.

Disruption of the mouse gene encoding the transcription factor MASH1 leads to loss of certain classes of neurons, including receptor neurons of the olfactory epithelium (OE). Here we investigate the nature of the cell type expressing MASH1 in mouse OE by manipulating olfactory receptor neuron (ORN) neurogenesis in vitro and in vivo to alter the dynamics of neuronal production. The results indicate that MASH1 is expressed in cells of the ORN lineage, but not in ORNs themselves nor in their immediate precursors. Data on how changes in the numbers and proliferative states of MASH+ cells correlate with induced changes in overall neurogenesis strongly suggest that MASH1-expressing cells give rise to the immediate precursors of ORNs, but are not the self-renewing stem cells of the OE. The results imply that multiple progenitor stages are employed in generating ORNs and suggest that the action of MASH1 occurs predominantly at an intermediate stage.

Animals↗

PubMed1995

Caffeine and smoking: subjective, performance, and psychophysiological effects.

The effects of caffeine and smoking on cognitive performance, subjective variables, heart rate, and EEG were assessed in two sessions. In one session, subjects received caffeine (2.5 mg/kg bodyweight), while in the other they received placebo. In both sessions they smoked a cigarette (8 cued puffs) having a nicotine yield of 1.2 mg. Caffeine produced an increase in self-reported muscular tension and tended to increase anxiety and delta magnitude. Smoking facilitated performance of a paper-and-pencil math task and increased heart rate. Smoking also appeared to produce cortical activation as indexed by decreased right frontal delta, decreased right centro-parietal theta, globally increased alpha, and increased centro-occipital/decreased posterior-temporal beta 1. Smoking also increased central/decreased posterior-temporal beta 2. Smoking and caffeine did not interact for any measure, suggesting that the epidemiological link between smoking and coffee drinking may have a non-pharmacological basis.

Adult↗